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目的:研究外源茉莉酸甲酯(Me JA)处理对中药何首乌(Polygonum multiflorum)根转录组的影响,分析在外源Me JA刺激下何首乌根各代谢途径中差异表达基因的变化,为寻找影响何首乌根主要药效成分合成的关键酶基因奠定基础。创新点:首次在中药何首乌上利用外源Me JA处理研究何首乌根转录组的变化,且获得了根中各代谢途径的大量差异表达基因。方法:将何首乌植株分为对照组(Hoagland营养液灌根)和实验组(含0.25 mmol/L MeJ A的Hoagland营养液灌根)。灌根处理26 h后,分别提取对照组和实验组根中的总RNA,利用新一代测序技术分析对照组和实验组根的转录组变化并进行测序。同时,利用美国国家生物技术信息中心(NCBI)的蛋白质数据库和基因数据库信息,对所测cD NA序列进行基因功能注释和代谢途径归类,寻找Me JA刺激下何首乌根各条代谢途径中的差异表达基因。结论:外源Me JA刺激处理可影响中药何首乌根转录组的表达变化(表S1)。与对照相比,外源Me JA处理可导致何首乌根中4535个基因上调,14 142个基因下调变化,并分别在125个代谢途径中富集(表1)。代谢途径差异表达基因分析的结果表明,可在相应代谢途径中找到影响何首乌某些主要药效成分合成的关键酶基因(图2)。差异表达基因的代谢途径归类分析结果提示今后可对何首乌根各代谢途径中的关键酶基因分别进行研究(表1)。综上所述,利用外源Me JA处理后进行转录组测序对寻找何首乌中主要有效成份合成的关键酶基因有重要价值。
OBJECTIVE: To study the effect of exogenous Me JA treatment on the root transcriptome of Polygonum multiflorum, and to analyze the changes of differentially expressed genes in the metabolic pathway of Polygonum multiflorum under the stimulation of exogenous Me JA. Root of the main pharmacodynamic components of the key enzyme genes laid the foundation. Innovation point: For the first time, the use of exogenous Me JA treatment of Polygonum multiflorum to study the changes of Radix Polygoni Multiflori root transcriptome, and obtained a large number of differentially expressed genes in the root metabolic pathway. Methods: Polygonum multiflorum plants were divided into control group (Hoagland nutrient solution irrigation) and experimental group (Hoagland nutrient solution containing 0.25 mmol / L MeJ A). After the roots were treated for 26 h, the total RNA in the roots of the control group and the experimental group were extracted, and the changes of the transcriptome of the control group and the experimental group were analyzed by a new generation of sequencing technology and sequenced. At the same time, the NCBI protein database and gene database information were used to classify the functional cDNA sequences annotated and metabolic pathways, in order to find out the differences among the metabolic pathways in the root of Polygonum multiflorum under Me JA stimulation Expression of genes. CONCLUSION: Exogenous Me JA stimulation can affect the expression of Transcription of Radix Polygoni multiflori (Table S1). Compared with the control, exogenous Me JA treatment resulted in up-regulation of 4535 genes in Polygonum multiflorum and 14,142 genes down-regulated, which were enriched in 125 metabolic pathways, respectively (Table 1). Metabolic pathway differentially expressed gene analysis results show that in the corresponding metabolic pathways to find the key enzyme genes that affect the synthesis of some of the main active ingredients of Polygonum multiflorum (Figure 2). Metabolic pathway of differentially expressed genes The classification analysis suggested that the key enzyme genes in each metabolic pathway of Polygonum multiflorum could be studied separately (Table 1). In summary, the use of exogenous Me JA treatment transcriptome sequencing to find the main active ingredients of Polygonum multiflorum synthesis of key enzyme genes of great value.